Immunogold Localization of Endothelin-1 in Endothelial Cells

摘要: Cultured endothelial cells secrete endothelin-1 (ET-1), a 21 amino-acid peptide, via the constitutive pathway of protein secretion. Constitutive secretory vesicle facilitate transport newly synthesized peptides from trans Golgi network to cell surface. In addition their role in intracellular peptide transport, serve as an important site for processing. ET-1 is processed its larger precursor molecule, Big ET-1, by proteolytic cleavage Trp21-Val22 bond. This unusal thought require novel protease which referred endothelin-converting-enzyme (ECE) (Yanagisawa et al., 1988). However this endopeptidase has proved technically difficult identify (Opgenorth 1992). As processing other prohormones occurs vesicles, we hypothesized that converted vesicles cells. study were isolated bovine aortic (BAEC) using discontinuous sucrose density gradient ultracentrifugation (Harrison 1993). The vesicle-containing fraction was identified specific [16–21] radioimmunoassay (Corder